WO2012109458A1 - Dérivés de flavine - Google Patents

Dérivés de flavine Download PDF

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Publication number
WO2012109458A1
WO2012109458A1 PCT/US2012/024507 US2012024507W WO2012109458A1 WO 2012109458 A1 WO2012109458 A1 WO 2012109458A1 US 2012024507 W US2012024507 W US 2012024507W WO 2012109458 A1 WO2012109458 A1 WO 2012109458A1
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Prior art keywords
alkyl
infection
methyl
compound
formula
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Philip D.G. Coish
Paul Adrian Aristoff
Brian R. Dixon
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BIORELIX Inc
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BIORELIX Inc
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Priority claimed from PCT/US2011/000617 external-priority patent/WO2011126567A1/fr
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/90—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having two or more relevant hetero rings, condensed among themselves or with a common carbocyclic ring system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/04—Ortho-condensed systems
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to novel flavin derivatives, composition comprising the same, and their use and compositions for use as riboswitch ligands and/or anti-infectives.
  • RNA structures termed riboswitches regulate the expression of various genes crucial for survival or virulence.
  • members of each known class of riboswitch can fold into a distinct, three-dimensionally structured receptor that recognizes a specific organic metabolite.
  • the riboswitch receptor binds to the metabolite and induces a structural change in the nascent mRNA that prevents expression of the open reading frame (ORF), thereby altering gene expression.
  • ORF open reading frame
  • the riboswitch folds into a structure that does not interfere with the expression of the ORF.
  • Riboswitch motifs have been identified that bind to thiamine pyrophosphate (TPP), flavin mononucleotide (FMN), glycine, guanine, 3'-5'-cyclic eiguanylic acid (c-di-GMP), molybdenum cofactor, glucosamine-6-phosphate (GlcN6P), lysine, adenine, and adocobalamin (AdoCbl) riboswitches.
  • TPP thiamine pyrophosphate
  • FMN flavin mononucleotide
  • c-di-GMP 3'-5'-cyclic eiguanylic acid
  • GlcN6P glucosamine-6-phosphate
  • AdoCbl adocobalamin
  • riboswitch motifs have been identified that recognize S-adenosylmethionine (SAM) I, II and III, IV and two distinct motifs that recognize pre-queosine- 1 (PreQl).
  • SAM S-adenosylmethionine
  • PreQl pre-queosine- 1
  • PTPP pyrithiamine pyrophosphate
  • AEC L- aminoethylcysteine
  • DL-4-oxalysine which bind to lysine riboswitches and roseoflavin and FMN which bind to FMN riboswitches.
  • the riboswitch-receptors bind to their respective ligands in an interface that approaches the level of complexity and selectivity of proteins. This highly specific interaction allows riboswitches to discriminate against most intimately related analogs of ligands.
  • the receptor of a guanine- binding riboswitch from Bacillus subtilis forms a three-dimensional structure such that the ligand is almost completely enveloped.
  • the guanine is positioned between two aromatic bases and each polar functional group of the guanine hydrogen bonds with four additional riboswitch nucleotides surrounding it. This level of specificity allows the riboswitch to discriminate against most closely related purine analogs.
  • SAM- binding riboswitches comprise one subdomain that recognizes every polar functional group of the 4-amino-5-hydroxymethyl-2- methylpyrimidine (HMP) moiety, albeit not the thiazole moiety, and another subdomain that coordinates two metal ions and several water molecules to bind the negatively charged pyrophosphate moiety of the ligand.
  • HMP 4-amino-5-hydroxymethyl-2- methylpyrimidine
  • FMN riboswitches Similar to TPP, guanine and SAM riboswitches, FMN riboswitches form receptor structures that are highly specific for the natural metabolite FMN. It is by this highly specific interaction that allows for the design of small molecules for the regulation of specific genes.
  • FMN riboswitches are of particular interest of this invention because it is believed that the riboswitch binds to flavin mono-nucleotide (FMN) and represses the expression of enzymes responsible for riboflavin and FMN biosynthesis.
  • Riboflavin is a water-soluble vitamin that is converted by flavokinases and FAD synthases to co-factors FMN and FAD, which are indispensable cofactors involved in energy metabolism and metabolism of fats, ketones, carbohydrates and proteins crucial for all living organisms.
  • flavokinases and FAD synthases are indispensable cofactors involved in energy metabolism and metabolism of fats, ketones, carbohydrates and proteins crucial for all living organisms.
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene) optionally substituted with one or more Ci- 6 alkyl (e.g., methyl) or one hydroxy or Ci
  • X is a single bond, -S- or -0-;
  • A is aryl (e.g., phenyl) or heteroaryl (e.g. pyridinyl) optionally substituted with one or more Ci ⁇ alkyl (e.g., methyl ) , Ci_ 4 alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi ⁇ alkyl (e g., CF 3 ) or -0-haloCi_ 4 alkyl (e.g.
  • Ri is H, Ci_ 4 alkyl (e.g., methyl, ethyl, n-propyl or isopropyl), or Ci_ 4 alkoxy (e.g., methoxy);
  • R 2 is H, Ci- 4 alkyl (e.g., methyl or n-propyl), Ci_ 4 alkoxy (e.g., methoxy), halo (e.g., CI), C 3 - 8 cycloalkyl-Ci_ 4 alkyl, -Ci ⁇ alkyl-N(R a )(R b ), (Ci ⁇ alkoxy)- Ci ⁇ alkyl or (2-Ci_ 4 alkoxyethoxy)-Ci ⁇ alkyl;
  • R3 is H or Ci ⁇ alkyl (e.g., methyl);
  • R 4 is H or Ci ⁇ alkyl (e.g., methyl);
  • R a and R b are independently H, Ci ⁇ alkyl (e.g., methyl) or C 3 _ 8 cycloalkyl (e.g., cyclopropyl, cyclopentyl);
  • the invention provides a compound of Formula IF ' :
  • Ci_ 6 alkylene e.g., n-propylene, n-butylene, n-pentylene
  • Ci_ 6 alkyl e.g., methyl
  • Ci_ 4alkoxy group e.g., methyl
  • ( ⁇ ) X is a single bond, -S- or -0-;
  • A is aryl (e.g., phenyl) or heteroaryl (e.g. pyridinyl) optionally substituted with one or more Ci ⁇ alkyl (e.g., methyl ) , Ci_ 4 alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi ⁇ alkyl (e . g., CF 3 ) or -0-haloCi_ 4 alkyl (e.g., -OCF 3 );
  • Ri is H or Ci ⁇ alkyl (e.g., methyl), or Ci ⁇ alkoxy (e.g., methoxy);
  • R 2 is H or Ci ⁇ alkyl (e.g., methyl), Ci_ 4 alkoxy (e.g., methoxy), halo (e.g.,
  • R 3 is H or Ci ⁇ alkyl (e.g., methyl);
  • R 4 is H or Ci ⁇ alkyl (e.g., methyl);
  • R a and 3 ⁇ 4 are independently H, Ci ⁇ alkyl (e.g., methyl) or C3_ 8 cycloalkyl (e.g., cyclopropyl, cyclopentyl);
  • the invention provides a compound of Formula II:
  • Ci_ 6 alkylene e.g., n-propylene, n-butylene, n-pentylene
  • Ci_ 4 alkoxy group optionally substituted with one hydroxy or Ci_ 4 alkoxy group
  • ( ⁇ ) X is a single bond, -S- or -0-;
  • A is aryl (e.g., phenyl) or heteroaryl (e.g. pyridinyl) optionally substituted with one or more Ci ⁇ alkyl (e.g., methyl), Ci ⁇ alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi ⁇ alkyl (e . g., CF 3 ) or -0-haloCi_ 4 alkyl (e.g., -OCF 3 );
  • Ri is H, Ci_ 4 alkyl (e.g., methyl) or Ci ⁇ alkoxy (e.g., methoxy);
  • R 2 is H, Ci_ 4 alkyl (e.g., methyl), Ci ⁇ alkoxy (e.g., methoxy), halo (e.g., CI), C 3 - 8 cycloalkyl-Ci ⁇ alkyl, -Ci_ 4 alkyl-N(R a )(R b ), (Ci_ 4 alkoxy)-Ci_ 4 alkyl or (2- Ci _ 4 alkoxyethoxy) -C 1 _ 4 alkyl ;
  • R3 is H or Ci ⁇ alkyl (e.g., methyl);
  • R 4 is H or Ci ⁇ alkyl (e.g., methyl);
  • R a and R b are independently H, Ci ⁇ alkyl (e.g., methyl) or C 3 _ 8 cycloalkyl (e.g., cyclopropyl, cyclopentyl);
  • the invention provides a compound of the following formulae: a compound of Formula P, wherein Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene) optionally substituted with one or more Ci_ 6 alkyl
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene) optionally substituted with one or more Ci_ 6 alkyl
  • Ci_ 4 alkoxy e.g., ethoxy
  • Alk is n-propylene substituted with one or more Ci_ 6 alkyl (e.g., methyl), for example Alk is
  • Ci_ 6 alkylene e.g., n-propylene, n-butylene, n-pentylene
  • Ci_ 6 alkylene e.g., n-propylene, n-butylene, n-pentylene
  • a compound of Formula P, IF ' or II or any of 2.1-2.6 wherein A is aryl (e.g., phenyl) or heteroaryl (e.g. pyridinyl) optionally substituted with one or more Ci_ 4 alkyl (e.g., methyl), Ci ⁇ alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi_ 4 alkyl (e.g., CF 3 ) or -O-haloCi ⁇ alkyl (e.g., -OCF 3 ); a compound of Formula P, IF ' or II or any of 2.1-2.7, wherein A is aryl (e.g., phenyl);
  • a compound of Formula P, IF ' or II or any of 2.1-2.8 wherein A is phenyl; 2.10 a compound of Formula P, IF ' or II or any of 2.1-2.7, wherein A is aryl (e.g., phenyl) substituted with one or more Ci- 4 alkyl (e.g., methyl) or halo (e.g., CI, F);
  • Ci ⁇ alkoxy e.g., methoxy
  • Ci ⁇ alkyl e.g., methyl
  • Ci ⁇ alkoxy e.g., methoxy
  • Ci ⁇ alkoxy e.g., methoxy
  • halo e.g., CI
  • Ci ⁇ alkyl e.g., methyl
  • Ci ⁇ alkoxy e.g., methoxy
  • halo e.g., CI
  • Ci ⁇ alkyl e.g., methyl
  • Ci ⁇ alkoxy e.g., methoxy
  • halo e.g., CI
  • Ci ⁇ alkyl e.g., methyl
  • Ci ⁇ alkyl e.g., methyl
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene)
  • Ci_ 6 alkyl e.g., methyl
  • Ci_ 4 alkoxy e.g., ethoxy
  • A is aryl (e.g., phenyl) optionally substituted with one or more C ⁇ alkyl (e.g., methyl) or halo (e.g., CI, F);
  • Ri is H or Ci_ 4 alkyl (e.g., methyl, ethyl, n-propyl or isopropyl);
  • R 2 is H or Ci_ 4 alkyl (e.g., methyl or n-propyl);
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene)
  • X is a single bond, -S- or -0-;
  • A is aryl (e.g., phenyl) optionally substituted with one or more Ci_ 4alkyl (e.g., methyl), Ci ⁇ alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi ⁇ alkyl (e.g., CF 3 ) or -O-haloCi ⁇ alkyl (e.g., -
  • Ri is Ci ⁇ alkyl (e.g., methyl);
  • R 2 is Ci ⁇ alkyl (e.g., methyl);
  • R 3 is H
  • R t is H
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene) optionally substituted with one hydroxy or Ci_ 4 alkoxy group;
  • X is a single bond;
  • A is aryl (e.g., phenyl) optionally substituted with one or more Ci_ 4alkyl (e.g., methyl), Ci ⁇ alkoxy (e.g., methoxy), hydroxy, halo (e.g., CI, F), haloCi ⁇ alkyl (e.g., CF 3 ) or -O-haloCi ⁇ alkyl (e.g., - OCF 3 );
  • Ci_ 4alkyl e.g., methyl
  • Ci ⁇ alkoxy e.g., methoxy
  • hydroxy halo
  • haloCi ⁇ alkyl e.g., CF 3
  • -O-haloCi ⁇ alkyl e.g., - OCF 3
  • Ri is Ci ⁇ alkyl (e.g., methyl);
  • R 2 is Ci ⁇ alkyl (e.g., methyl);
  • R 3 is H
  • Alk is Ci_ 6 alkylene (e.g., n-propylene, n-butylene, n-pentylene); X is a single bond;
  • A is aryl (e.g., phenyl);
  • Ri is Ci ⁇ alkyl (e.g., methyl);
  • R 2 is Ci ⁇ alkyl (e.g., methyl);
  • R 3 is H
  • MIC Minimum Inhibitory Concentration
  • the compound of the invention is a compound as hereinbefore described (e.g., Formula P, II", IF, II, or any of 2.1-2.31), wherein X is a single bond and the remaining substituent is as defined in any of the formula described therein (e.g., Formula P, ⁇ ", IF , II, or any of 2.1-2.31), in free or salt form.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula P, II" or II, e.g., any of formulae 2.1-2.31 , in free or pharmaceutically acceptable salt form in admixture with a pharmaceutically acceptable diluent or carrier.
  • the invention provides a method for the treatment or prophylaxis of a bacterial infection (Method P) comprising administering to a subject in need thereof an effective amount of a compound of Formula P, IF ' or II, e.g., any of formulae 2.1-2.31, in free or pharmaceutically acceptable salt form.
  • Methods P as hereinbefore described are useful for the treatment or prophylaxis of a Gram-positive or Gram- negative bacterial infection (Method P-A).
  • Method P is useful for treating a bacterial infection including, but not limited to an infection by one or more of the following bacteria: Clostridium difficile (or C.
  • Method P is particularly useful for treating an infection caused by Clostridium difficile.
  • the Compounds of the Invention, particularly Examples 1-16 are also active against the FMN riboswitch.
  • Compounds which are active against FMN riboswitch are generally also active against Staphylococcus aureus and/or Clostridium difficile infections.
  • the compounds of the invention may also be useful for the treatment of a Staphylococcus aureus infection.
  • Method P as hereinbefore described is useful for the treatment or prophylaxis of a disease, infection or condition selected from a group consisting of anthrax, staphylococcal scalded skin syndrome (staph infections), pneumonia, impetigo, boils, cellulitis, folliculitis, furuncles, carbuncles, scalded skin syndrome, abscesses, meningitis, osteomyelitis endocarditis, Toxic Shock Syndrome (TSS), septicemia, acute sinusitis, otitis media, septic arthritis, endocarditis, peritonitis, pericarditis, brain abscess, tularemia, urinary tract infection, empyema, food poisoning, diarrhea, conjunctivitis and Clostridium difficile associated disease (CDAD), comprising administering to a subject in need thereof an effective amount of a compound of Formula P, II" or II, e.g.,
  • the current invention provides methods of treating a bacterial infection via a novel mechanism, e.g., by utilizing riboswitch-ligand binding to alter gene expression. Therefore in one aspect, the compounds of the invention, particularly compounds of Examples 1-16 bind to FMN riboswitches, thereby affecting downstream riboflavin biosynthesis. In another aspect, various compounds of the invention are also active against the CD3299 riboswitch, thereby affecting expression of the adjacent coding region. Compounds that are active against CD3299 and/or FMN riboswitch are particularly selective against C. difficile.
  • the Compounds of the Invention e.g., the compounds of Formula P, II" or II, e.g., any of formulae 2.1-2.31, particularly Examples 1-16, in free or pharmaceutically acceptable salt form, are effective in treating an infection wherein traditional antibiotics are rendered ineffective due to drug resistance. Therefore, in a particular embodiment, the invention provides Method P as hereinbefore described wherein the infection is by an infectious agent which is resistant to a drug that is not a riboswitch ligand (Method P-E). In a further embodiment, the infection to be treated in Method P is a C.
  • the infection is by an infectious agent which is resistant to any drug that is not a riboswitch ligand, e.g., fluoroquinolone (e.g., ciprofloxacin- and/or levofloxacin-resistant infection), metronidazole and/or vancomycin.
  • the compounds of Formula P, ⁇ " or II e.g., any of formulae 2.1-2.31, in free or pharmaceutically acceptable salt form may be useful for a Staphylococcus aureus infection which is resistant to one or more drugs selected from a group consisting of a penicillin, vancomycin, cephalosporin and methicillin.
  • the infection is a methicillin-resistant Staphylococcus aureus infection.
  • various compounds of the Invention have a low CC5 0 value in an assay as disclosed in Example C and therefore, may have anti-metabolite activities which may interfere with DNA biosynthesis. Therefore, in one embodiment, these compounds may be useful as an anti-cancer or anti- viral agent. In another embodiment, the compounds that have a low MIC and/or a high I max value in an assay as disclosed in Example B and A respectively, and a low CC5 0 value in an assay as disclosed in Example C are used as an antibacterial, for topical administration.
  • the invention provides a method for the treatment of an infection in a plant (Method P-F) comprising administering to such plant an effective amount of a compound of Formula P, II" or II, e.g., any of formulae 2.1-2.31 , in free or pharmaceutically acceptable salt form.
  • the invention provides use of a compound, or use of a pharmaceutical composition comprising a compound of Formula P, II" or II, e.g., any of formulae 2.1-2.31, in free or pharmaceutically acceptable salt form, (in the manufacture of a medicament) for the treatment or prophylaxis of an infection, e.g., a bacterial infection as described in Methods P, or any of Methods P-A through P-F.
  • a compound of Formula P, II" or II e.g., any of formulae 2.1-2.31, in free or pharmaceutically acceptable salt form, (in the manufacture of a medicament) for the treatment or prophylaxis of an infection, e.g., a bacterial infection as described in Methods P, or any of Methods P-A through P-F.
  • the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of Formula P, IF ' or II, e.g., any of formulae 2.1-2.31 , in free or pharmaceutically acceptable salt form, for use in the treatment of any disease or condition as hereinbefore described, e.g., in any of Methods P or Methods P-A through P-F.
  • riboswitch or "riboswitches” is an art recognized term and refers to an mRNA which comprises a natural aptamer that binds target metabolite and an expression platform which changes in the RNA structure to regulate genes.
  • riboswitch ligand refers to any compound such as a compound of Formula P, II” or II, e.g., any of formulae 2.1-2.31 ,, in free or salt form, that binds to that particular riboswitch.
  • FMN riboswitch refers to a riboswitch that binds a metabolite such as flavin mono-nucleotide (FMN) or other ligands such as the compound of Formula P, II" or II, e.g., any of formulae 2.1-2.31, in free or salt form, and which affects downstream FMN biosynthesis and transport proteins.
  • FMN flavin mono-nucleotide
  • the binding of the ligand to its riboswitch induces a conformational change in the bacterial mRNA such that the expression of the ORF is altered, for example, such that the expression of enzymes responsible for, e.g., riboflavin and FMN biosynthesis is repressed or overexpressed.
  • CD3299 riboswitch refers to a riboswitch found in C. difficile, controlling the gene designated CD3299.
  • accession number AM180355 is as follows:
  • ORF start site in the above sequence is downstream from the riboswitch and is depicted in italics and is:
  • the putative terminator hairpin is in bold italics and is:
  • the hairpin can form a loop having a structure as depicted in Formula 1 :
  • a possible antiterminator has a structure as depicted in Formula 2:
  • infection encompasses an infection by a Gram-positive or Gram- negative bacteria.
  • the infection is by a Gram-positive bacteria.
  • the infection is by a Gram-negative bacteria.
  • the infection is an infection by one or more bacteria selected from a group consisting of Clostridium difficile, Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii,
  • Escherichia coli Haemophilus influenzae, Enterococcus faecalis, Streptococcus pyogenes, Listeria monocytogenes, Salmonella enterica, Vibrio cholerae, Brucella melitensis, Bacillus anthracis, Francisella tularensis, Moraxella catarrhalis, Klebsiella pneumoniae, Yersinia pestis, Streptococcus viridans, Enterococcus faecium, and/or Borrelia burgdorferi.
  • the infection is a Clostridium difficile and/or Staphylococcus aureus infection.
  • the infection is an infection which is resistant to a drug which is not a riboswitch ligand.
  • the infection is an infection which is resistant to one or more drugs selected from a group consisting of penicillin, vancomycin, cephalosporin, methicillin and fluoroquinolone (e.g., ciprofloxacin- and/or levofloxacin).
  • the infection is a methicillin-resistant Staphylococcus aureus (MRSA) infection.
  • the infection is a fluoroquinolone-resistant (e.g., ciprofloxacin- and/or levofloxacin-resistant), metronidazole and/or vancomycin-resistant C. difficile infection.
  • bacteria or "bacterial” include, but are not limited to Clostridium difficile, Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Haemophilus influenzae, Enterococcus faecalis, Streptococcus pyogenes, Listeria monocytogenes,
  • Salmonella enterica Vibrio cholerae, Brucella melitensis, Bacillus anthracis, Francisella tularensis, Moraxella catarrhalis, Klebsiella pneumoniae, Yersinia pestis, Streptococcus viridans, Enterococcus faecium, and/or Borre lia burgdorferi .
  • Alkyl as used herein is a saturated or unsaturated hydrocarbon moiety, preferably saturated, e.g., one to eight, e.g., one to six, e.g., one to four carbon atoms in length, which may be linear or branched (e.g., n-butyl or tert-butyl) unless otherwise specified, and may be optionally substituted, e.g., mono-, di-, or tri-substituted on any one of the carbon atoms, e.g., with Ci_ 4 alkyl (e.g., methyl), Ci- 4 alkoxy, halogen (e.g., chloro or fluoro), haloCi ⁇ alkyl (e.g., trifluoromethyl), hydroxy, and carboxy.
  • Ci_ 4 alkyl e.g., methyl
  • Ci- 4 alkoxy e.g., halogen (e.g., chloro or fluoro), halo
  • C1-C8 alkyl denotes alkyl having 1 to 8 carbon atoms.
  • alkyl include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i- butyl, sec -butyl, t-butyl, 3-methylpentyl, 4-methylpentyl, n-pentyl, n-hexyl and n-heptyl.
  • alkylene is intended to denote an alkyl group bridging between two substituents (e.g., between the flavin core structure and another substituent, for example -X-A). Therefore Ci_ 4alkylene, e.g., methylene, ethylene, n-propylene and n-butylene are intended to represent -CH 2 - -CH 2 CH 2 -, -CH 2 CH 2 CH 2 - and - CH 2 CH 2 CH 2 CH 2 - respectively. Wherein the alkylene group is unsaturated or partially saturated, it is denoted as "alkenylene" or "alkynylene".
  • Aryl as used herein is a monocyclic or polycyclic aromatic hydrocarbon, preferably phenyl, optionally substituted, e.g., with Ci_ 4 alkyl (e.g., methyl), Ci ⁇ alkoxy, halogen (e.g., chloro or fluoro), haloCi ⁇ alkyl (e.g., trifluoromethyl), hydroxy, carboxy, or an additional aryl or heteroaryl.
  • Ci_ 4 alkyl e.g., methyl
  • Ci ⁇ alkoxy e.g., methyl
  • halogen e.g., chloro or fluoro
  • haloCi ⁇ alkyl e.g., trifluoromethyl
  • Heterocycloalkyl refers to a cycloalkyl as defined above wherein at least one of the carbon atoms is replaced with a heteroatom selected from N, O, S. Therefore, “C3_ 8 heterocycloalkyl” or “heteroC 3 _ 8 cycloalkyl” refers to a 3- to 8-membered non-aromatic ring system containing at least one heteroatom selected from N, O and S.
  • the substituent is connected via an alkyl group, e.g., -Co- 4 alkyl-C 3 _ gcycloalkyl or aryl-Ci ⁇ alkyl, it is understood that the alkyl group may be saturated or unsaturated or linear or branched.
  • the substituent is connected via the Co-alkyl, it is understood that the alkyl is not present and the connectivity is directly to the next substituent.
  • the substituent is -Coalkyl-C3_ 8 cycloalkyl, it is understood that the alkyl group is not present and the cycloalkyl (e.g., cyclopropyl) is directly connected.
  • An acid-addition salt of a compound of the invention which is sufficiently basic for example, an acid-addition salt with, for example, an inorganic or organic acid, for example hydrochloric, hydrobromic, sulphuric, phosphoric, acid acetic, trifluoroacetic, citric, maleic acid, toluene sulfonic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, hydroxy maleic, phenylacetic, glutamic, benzoic, salicylic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disul
  • an inorganic or organic acid for example hydrochloric, hydrobromic, sulphuric, phosphoric, acid acetic, trifluoroacetic, citric, maleic acid, toluene sulfonic
  • a salt of a compound of the invention which is sufficiently acidic is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium or magnesium salt, an ammonium salt or a salt with an organic base which affords a physiologically-acceptable cation, for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.
  • the salt of the compound of the invention is a trifluoroacetic or hydrochloric acid addition salt.
  • the salt of the compound of the invention is an acetic acid addition salt.
  • Compounds of the Invention is to be understood as embracing the compounds disclosed herein, such as a compound of Formula P, II" or II, e.g., any of formulae 2.1 -2.31 , in any form, for example free or acid addition salt or prodrug form, or where the compounds contain acidic substituents, in base addition salt form.
  • the Compounds of the Invention are intended for use as pharmaceuticals, therefore pharmaceutically acceptable salts are preferred. Salts which are unsuitable for pharmaceutical uses may be useful, for example, for the isolation or purification of free Compounds of the Invention, and are therefore also included.
  • the Compounds of the Invention may comprise one or more chiral carbon atoms.
  • the compounds thus exist in individual isomeric, e.g., enantiomeric or diasteriomeric form or as mixtures of individual forms, e.g., racemic/diastereomeric mixtures. Any isomer may be present in which the asymmetric center is in the (R)-, (S)-, or (R,S)- configuration.
  • the invention is to be understood as embracing both individual optically active isomers as well as mixtures (e.g., racemic/diasteromeric mixtures) thereof.
  • the Compound of the Invention may be a racemic mixture or it may be predominantly, e.g., in pure, or substantially pure, isomeric form, e.g., greater than 70% enantiomeric excess ("ee"), preferably greater than 80% ee, more preferably greater than 90% ee, most preferably greater than 95% ee.
  • ee enantiomeric excess
  • the purification of said isomers and the separation of said isomeric mixtures may be accomplished by standard techniques known in the art (e.g., column chromatography, preparative TLC, preparative HPLC, simulated moving bed and the like).
  • the Compounds of the Invention encompass their stable isotopes.
  • the hydrogen atom at a certain position on the Compounds of the Invention may be replaced with deuterium. It is expected that the activity of compounds comprising such isotopes would be retained and/or it may have altered pharmacokinetic or pharmacodynamic properties.
  • pharmacodynamic properties would also have utility for measuring pharmacokinetics of the non-isotopic analogs.
  • Compounds of the Invention may in some cases also exist in prodrug form.
  • prodrug is an art recognized term and refers to a drug precursors prior to administration, but generate or release the active metabolite in vivo following
  • the Compounds of the Invention e.g., a compound of Formula P, IF ' or II, e.g., any of formulae 2.1-2.31
  • these substituents may be esterified to form physiologically hydrolysable and acceptable esters (e.g., acyl esters, e.g., CH 3 C(0)-0- Compound).
  • physiologically hydrolysable and acceptable esters means esters of Compounds of the Invention which are hydrolysable under physiological conditions to yield hydroxy on the one hand and acid, e.g., carboxylic acid on the other (e.g., Drug-0-C(0)-CH 3 -» Drug-OH + CH 3 COOH), which are themselves
  • amide prodrugs may also exist wherein the prodrug is cleaved to release the active amine metabolite in vivo following administration. Further details of amine prodrugs may be found in Jeffrey P. Krise and Reza Oliyai, Biotechnology: Pharmaceutical Aspects, Prodrugs, Volume 5, Part 3, pages 801-831, the contents of which are herein incorporated by reference in their entirety. As will be appreciated, the term thus embraces conventional pharmaceutical prodrug forms.
  • the Compounds of the Invention are useful for the treatment of an infection, particularly an infection by bacteria including but not limited to Clostridium difficile, Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Haemophilus influenzae, Enterococcus faecalis, Streptococcus pyogenes, Listeria monocytogenes, Salmonella enterica, Vibrio cholerae, Brucella melitensis, Bacillus anthracis, Francisella tularensis, Moraxella catarrhalis, Klebsiella pneumoniae, Yersinia pestis, Streptococcus viridans, Enterococcus faecium, and/or Borrelia burgdorferi bacteria.
  • the bacteria is Clostridium difficile.
  • the invention therefore provides methods of treatment of any one or more of the following conditions: anthrax infection, staphylococcal scalded skin syndrome (staph infections), pneumonia, impetigo, boils, cellulitis, folliculitis, furuncles, carbuncles, scalded skin syndrome, abscesses, meningitis, osteomyelitis endocarditis, Toxic Shock Syndrome (TSS), septicemia, acute sinusitis, otitis media, septic arthritis, endocarditis, peritonitis, pericarditis, brain abscess, tularemia, urinary tract infection, empyema, food poisoning, diarrhea, conjunctivitis and Clostridium difficile associated disease (CD AD); comprising administering an effective amount of a compound of Formula P, II" or II, e.g., any of formulae 2.1-2.31, in free or pharmaceutically acceptable salt form, to a subject in need thereof.
  • treatment and “treating” are to be understood accordingly as embracing prophylaxis and treatment or amelioration of symptoms of disease as well as treatment of the cause of the disease.
  • the invention encompasses prophylaxis of symptoms of disease or cause of the disease.
  • the invention encompasses treatment or amelioration of symptoms of disease or cause of the disease.
  • subject encompasses human and/or non-human
  • a therapeutically active amount of the therapeutic compositions is defined as an amount effective, at dosages and for periods of time necessary to achieve the desired result.
  • a therapeutically effective amount of a Compound of the Invention reactive with at least a portion of the FMN or the CD3299 riboswitch may vary according to factors such as the disease state, age, sex, and weight of the individual, and the ability of the compound to elicit a desired response in the individual.
  • Dosage regiment may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. In general, satisfactory results, e.g.
  • Unit dosage forms for oral administration thus for example may comprise from about 0.2 to 75 mg, 250 mg,1000 mg, e.g. from about 0.2 or 2.0 to 50, 75, 100, 250, 500, 750 or 1000 mg of a Compound of the Invention, together with a pharmaceutically acceptable diluent or carrier therefor.
  • compositions comprising the Compounds of the Invention may be prepared using conventional diluents or excipients and techniques known in the galenic art.
  • oral dosage forms may include tablets, capsules, solutions, suspensions, spray-dried dispersions [e.g. Eudragit L100] and the like.
  • pharmaceutically acceptable carrier as used herein is intended to include diluents such as saline and aqueous buffer solutions.
  • the Compounds of the Invention may be administered in a convenient manner such as by injection such as subcutaneous, intravenous, by oral administration, inhalation, transdermal application, intravaginal application, topical application, intranasal, sublingual or rectal administration.
  • the active compound may be coated in a material to protect the compound from the degradation by enzymes, acids and other natural conditions that may inactivate the compound.
  • the compound may be orally administered.
  • the compound is administered via topical application.
  • the Compounds of the Invention may be administered alone or in conjunction, e.g., at or about the same time or simultaneously and separately or simultaneously in an admixture, with another agent, e.g., an agent to facilitate entry or permeability of the Compounds of the Invention into the cell, e.g., an antimicrobial cationic peptide.
  • Antimicrobial cationic peptides include peptides which contain (1) a disulfide-bonded ⁇ -sheet peptides; (2) amphipathic a-helical peptides; (3) extended peptides; or (4) loop-structured peptides.
  • cationic peptide examples include but are not limited to defensins, cecropins, melittins, magainins, indolicidins, bactenecin and protegrins.
  • antimicrobial cationic peptides include but are not limited to human neutrophil defensin-1 (HNP-1), platelet microbicidal protein- 1 (tPMP), inhibitors of DNA gyrase or protein synthesis, CP26, CP29, CP11CN, CPIOA, Bac2A- NH 2 as disclosed in Friedrich et al., Antimicrob. Agents Chemother. (2000) 44(8):2086, the contents of which are hereby incorporated by reference in its entirety.
  • Further examples of antibacterial cationic peptides include but are not limited to polymyxin e.g., polymixin B, polymyxin E or polymyxin nonapeptide. Therefore, in another embodiment, the
  • Compounds of the Invention may be administered in conjunction with polymyxin, e.g., polymyxin B, polymyxin E or polymyxin nonapeptide, preferably polymyxin B.
  • polymyxin e.g., polymyxin B, polymyxin E or polymyxin nonapeptide, preferably polymyxin B.
  • the Compounds of the Invention may be administered alone or in conjunction, e.g., at or about the same time, simultaneously and separately, or simultaneously in an admixture, with other antimicrobial agents, e.g., other antifungal or other systemic antibacterial (bactericidal or bacteriostatic) agents.
  • other antimicrobial agents e.g., other antifungal or other systemic antibacterial (bactericidal or bacteriostatic) agents.
  • bacterial agents include agents which inhibit bacterial cell wall synthesis (e.g., penicillins, cephalosporins, carbapenems, vancomycin), agents which damage cytoplasmic membrane (e.g., polymixins as discussed above), agents which modify the synthesis or metabolism of nucleic acids (e.g., quinolones, rifampin, nitrofurantoin), agents which inhibit protein synthesis (aminoglycosides, tetracyclines, chloramphenicol, erythomycin, clindamycin), agents which interfer with the folate synthesis (e.g., folate-inhibitors), agents which modify energy metabolism (e.g., sulfonamides, trimethoprim) and/or other antibiotics (beta-lactam antibiotic, beta-lactamase inhibitors).
  • agents which inhibit bacterial cell wall synthesis e.g., penicillins, cephalosporins, carbapenems, vancomycin
  • the compounds of the Invention e.g., compound of Formula P, IF ' or II, e.g., any of formulae 2.1-2.31 , in free or salt form may be made using the methods as described and exemplified herein and by methods similar thereto and by methods known in the chemical art. Such methods include, but not limited to, those described below.
  • synthetic methods include, but not limited to, those described below.
  • all proposed reaction conditions including choice of solvent, reaction atmosphere, reaction temperature, duration of the experiment and workup procedures, are chosen to be the conditions standard for that reaction, which should be readily recognized by one skilled in the art. Therefore, at times, the reaction may require to be run at elevated temperature or for a longer or shorter period of time.
  • 2.1-2.31 may be prepared by reacting Intermediate-5 (Int-5) with ammonia in a pressure tube.
  • Int-5 may be prepared by reacting Intermediate-4 (Int-4) with diethyl 2-bromo-3- oxopentanedioate in the presence of a base, e.g., cesium carbonate, in a solvent, for example, a mixture of dimethylformamide (DMF) and methylene chloride (CH 2 CI 2 ).
  • a base e.g., cesium carbonate
  • Int- 4 may be may be prepared by converting Intermediate-3 (Int-3) to Int-4, for example, by catalytic hydrogenation, e.g., by reacting Int-3 with a metal, e.g., Raney-Nickel, and hydrogen gas in a solvent such as ethanol.
  • Int-3 may be prepared by reacting Intermediate-1 (Int-1) with NH 2 -Alk-X-A (Int-2), wherein Alk, X and A are defined in Formula P, IF ' or II or any of 2.1-2.31 to yield Int-3.
  • Int-1 is either commercially available or may be prepared by methods known in the art.
  • R 2 of compounds of Formula P, IF ' or II is alkoxy
  • this compound may be prepared by reacting a compound of Formula P, IF ' or II, wherein R 2 is halo, e.g., chloro, with R 2 -H, e.g., methanol, in the presence of a base.
  • R 2 is halo, e.g., chloro
  • R 2 -H e.g., methanol
  • R 2 of the compounds of Formula P, IF ' or II is (Ci ⁇ alkoxy)- methyl
  • these compounds may be prepared by first halogenating the compound of Formula P, IF ' or II, wherein R 2 is methyl, for example by reacting such compound with a halogen, e.g., bromine, e.g., optionally in the presence of a catalyst such as
  • azobisisobutyronitrile AIBN
  • the resulting intermediate, Int-6 may then react with a R 2 -H, wherein R 2 -H is e.g. methanol, in the presence of a base to provide the
  • R 2 of the compounds of Formula P, IF ' or II is -methyl-N(R a )(Rb), e.g., -CH 2 -N(CH 3 ) 2
  • this compound may be prepared by halogenating the compounds of Formula P, IF ' or II, wherein R 2 is e.g., a methyl group, for example by reacting bromine with the compounds of Formula P, IF ' or II, wherein R 2 is methyl, optionally in the presence of a catalyst such as azobisisobutyronitrile (AIBN).
  • AIBN azobisisobutyronitrile
  • the resulting intermediate, Int-6 may then react with an amine, HN(R a )(Rb), e.g.
  • R 2 of the compounds of Formula P, II" or II is hydrogen
  • these compounds may be prepared by heating Intermediate- 10 (Int-10) with acid [e.g.
  • Int-10 may be prepared by heating Intermediate-9 (Int- 9) with sodium dithionite in a mixture of ethanol and water.
  • Intermediate-9 (Int-9) may be prepared by treating Intermediate-8 (Int-8) with sodium nitrite in acetic acid with cooling.
  • Int-8 may be prepared by condensing Intermediate-7 (Int-7) with methyl 5-[(4- methoxybenzyl)amino]-3,5-dioxopentanoate in the presence of acid [e.g. p-toluenesulfonic acid monohydrate] in toluene.
  • Intermediate-7 (Int-7) may be prepared with Br- Alk-X-Ar , wherein Alk, X, and A are defined in any of Formulae P, II", or II. This preparation may be summarized in the following reaction scheme:
  • Precursor mRNA leader molecules are prepared by in vitro transcription from templates generated by PCR and [5'- 32 P] -labeling using methods described previously (Regulski and Breaker, In-line probing analysis of riboswitches (2008), Methods in Molecular Biology Vol 419, pp 53-67).
  • RNA precursor Approximately 5 nM of labeled RNA precursor is incubated for 41 hours at 25°C in 20 mM MgCl 2 , 50 mM Tris/HCl (pH 8.3 at 25°C) in the presence or absence of a fixed concentration of each ligand. Binding to the FMN and CD3299 riboswitches are measured at 20 ⁇ and 100 ⁇ , respectively. In-line cleavage products are separated on 10% polyacrylamide gel electrophoresis (PAGE), and the resulting gel is visualized using a Molecular Dynamics Phosphorimager. The location of products bands corresponding to cleavage are identified by comparison to a partial digest of the RNA with RNase Tl (G- specific cleavage) or alkali (nonspecific cleavage).
  • RNA In-line probing exploits the natural ability of RNA to self-cleave at elevated pH and metal ion concentrations (pH ⁇ 8.3, 25 mM MgCl 2 ) in a conformation-dependent manner.
  • the 2'-hydroxyl of the ribose For self-cleavage to occur, the 2'-hydroxyl of the ribose must be "in-line" with the phosphate-oxygen bond of the internucleotide linkage, facilitating a S 2P nucleophilic transesterification and strand cleavage.
  • single-stranded regions of the riboswitch are dynamic in the absence of an active ligand, and the internucleotide linkages in these regions can frequently access the required in-line conformation.
  • Binding of an active ligand to the riboswitch generally reduces the dynamics of these regions, thereby reducing the accessibility to the in-line conformation, resulting in fewer in-line cleavage events within those regions.
  • These ligand-dependent changes in RNA cleavage can be readily detected by denaturing gel electrophoresis.
  • the relative binding affinity of each ligand is expressed as I max , wherein I max represents the percent inhibition of in-line cleavage at selected internucleotide ligands in the presence of a fixed ligand concentration (20 ⁇ for the FMN riboswitch and 100 ⁇ for the CD3299 riboswitch) normalized to the percent inhibition in the absence of ligand and the percent inhibition in the presence of a saturation concentration of a control ligand.
  • 100 ⁇ FMN is used as a control ligand for estimating binding to the FMN riboswitch and 100 ⁇ of 7,8-dimethyl-10-(3- phenylpropyl)benzo[g]pteridine-2,4(3H,10H)-dione (which is a compound which has a high affinity against the CD3299 riboswitch) is used as a control ligand for estimating binding to the CD3299 riboswitch.
  • Example B [0048] The MIC assays are carried out in a final volume of 100 in 96-well clear round-bottom plates according to methods established by the Clinical Laboratory
  • test compound suspended in 100 % DMSO or another suitable solubilizing buffer
  • 100 % DMSO or another suitable solubilizing buffer
  • This solution is serially diluted by 2-fold into successive tubes of the same media to give a range of test compound concentrations appropriate to the assay.
  • 50 ⁇ of a bacterial suspension from an overnight culture growth in media appropriate to a given pathogen is added 50 ⁇ of a bacterial suspension from an overnight culture growth in media appropriate to a given pathogen.
  • Final bacterial inoculum is approximately 10 5 -10 6 CFU/well.
  • the MIC is defined as the lowest concentration of antimicrobial agent that completely inhibits growth of the organism as detected by the unaided eye, relative to control for bacterial growth in the absence of added antibiotic. Ciprofloxacin is used as an antibiotic-positive control in each screening assay.
  • Each of the bacterial cultures that are available from the American Type Culture Collection (ATCC, www.atcc.org) is identified by its ATCC number.
  • the exemplified compounds of the invention e.g., the compounds of Examples 1-16 have a minimum inhibitory concentration (MIC) of less than 2 ⁇ g/mL against at least one of the bacteria selected from C. difficile MMX3581 (clinical), C. difficile ATCC 43596, C. difficile ATCC 700057 (MMX 4381), C. difficile ATCC BAA-1805 (NAP1), C. difficile ATCC BAA-1382 (MMX4820), C. difficile 43255 (MMX4821), C. difficile ATCC BAA-1803 (NAP1) and C. difficile ATCC BAA-1870 (NAP1).
  • MIC minimum inhibitory concentration
  • All of the exemplified compounds of the invention have either an I max value of greater than 20% in an assay as described in Example A (compared to at least one of the two controls) and/or a MIC of less than or equal to 2 ⁇ g/mL against at least one of the bacterial strains as decribed in Example B.
  • certain compounds of the invention have either an I max value of greater than 50% in an assay as described in Example A (compared to at least one of the two controls) and/or a MIC of less than or equal to 2 ⁇ g/mL against at least one of the bacterial strains as decribed in Example B.
  • Example C The cytotoxic effects of test compounds on HepG2 are measured with a commercially available cell viability assay kit from Promega. On day 1, HepG2 cells ( ⁇ 1 x 10 4 cells) are seeded into each well in 96-well plate and cultured for approximately 24 h at 37°C in a 5% C0 2 atmosphere under saturating humidity. On day 2, test compounds and DMSO controls are added to appropriate wells to give a range of test compound concentrations appropriate to the assay. Terfenadine is also added to each plate as a positive cytotoxic control. Control wells containing medium without cell are prepared to obtain a value for background luminescence.
  • Assay plates are then cultured for approximately 24 h at 37°C in a 5% C02 atmosphere under saturating humidity. On day 3, assay plates are removed from 37°C incubator and equilibrated to 22°C. Once equilibrated, CellTiter-Glo ® reagent is added to each well containing cell culture medium, followed by mixing to allow cell lysis. The CellTiter-Glo ® Assay measures the number of viable cells in culture based on quantitation of the ATP present, an indicator of metabolically active cells. This assay generates a luminescent signal proportional to the amount of ATP present. The amount of ATP is directly proportional to the number of cells present in culture. After the assay plate is incubated at room temperature for
  • CC5 0 is defined as the concentration of test compounds in ⁇ to result in 50% reduction in luminescence signal relative to the signal for untreated cells.
  • the experiments show that the exemplified compounds of the invention have a CC5 0 value of greater than or equal to 19 ⁇ .
  • the exemplified compounds of the invention generally have a MIC to cytotox ratio of at least 1:20.
  • Method D Agilent 1100 HPLC, Agilent XDB C18 50 x 4.6 mm 1.8 micron column, 1.5 mL/min., Solvent A-Water (0.1% TFA), Solvent B -Acetonitrile (0.07% TFA), Gradient -5 min. 95%A to 95%B; lmin. hold; then recycle, UV Detection @ 210 and 254nm.
  • Method G Agilent 1100 HPLC, Agilent XDB C18 50 x 4.6 mm 5 micron column, 1.5 mL/min., Solvent A- Water (0.1% TFA), Solvent B -Acetonitrile (0.07% TFA), Gradient -6 min. 95%A to 95%B; 1 min. hold; then recycle, UV Detection @ 210 and 250nm.
  • NaHCC sodium bicarbonate
  • Na 2 S04 sodium sulfate
  • NH 3 ammonia gas
  • RNA ribonucleic acid
  • RNase Tl an endoribonuc lease that specifically degrades single-stranded RNA at G residues
  • 3,4-dihydroquinoxaline-2-carboxylate (0.300 g, 0.690 mmol) is taken up in 20 mL of MeOH and the solution is cooled in an ice water bath. Ammonia gas is bubbled through the solution for 5 minutes in a pressure tube. The solution is stirred at rt overnight in the capped pressure tube. The pressure tube is opened slowly to allow NH 3 to evolve. The remaining solution is evaporated to give 0.2 g of a dark solid. The solid is adsorbed onto silica gel and chromatographed on 50 g of silica gel. The column is eluted with 1% MeOH

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Abstract

La présente invention concerne de nouveaux dérivés de flavine, une composition les comportant, leur utilisation et des compositions à utiliser en tant que ligands riborégulateurs et/ou produits anti-infectieux.
PCT/US2012/024507 2011-02-09 2012-02-09 Dérivés de flavine Ceased WO2012109458A1 (fr)

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US201161441160P 2011-02-09 2011-02-09
US61/441,160 2011-02-09
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PCT/US2011/000617 WO2011126567A1 (fr) 2010-04-06 2011-04-06 Dérivés de flavine
US61/516,651 2011-04-06
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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3920650A (en) * 1973-09-19 1975-11-18 Morton Norwich Products Inc Isoalloxazines
WO2010019208A1 (fr) * 2008-08-11 2010-02-18 Biorelix Pharmaceuticals, Inc. Dérivés de la flavine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3920650A (en) * 1973-09-19 1975-11-18 Morton Norwich Products Inc Isoalloxazines
WO2010019208A1 (fr) * 2008-08-11 2010-02-18 Biorelix Pharmaceuticals, Inc. Dérivés de la flavine

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CARLSON ET AL.: "Improved Chemical Syntheses of 1- and 5-Deazariboflavin", J. ORG. CHEM., vol. 69, 2004, pages 2614 - 2617 *
KITTLEMAN ET AL.: "Characterization and Mechanistic Studies of Type II Isopentenyl Diphosphate: Dimethylallyl Diphosphate Isomerase from Staphylococcus aureus.", BIOCHEMISTRY, vol. 46, no. 28, 2007, pages 8401 - 8413, Retrieved from the Internet <URL:http://www.ncbi.nlm.nih.gov/pmGarticIes/PMC2515275/pdf/nihms60389.pdf> *
OTT ET AL.: "The RFN riboswitch of Bacillus subtilis is a target for the antibiotic roseoflavin produced by Streptomyces davawensis.", RNA BIOLOGY, vol. 6, no. 3, 2009, pages 276 - 280, Retrieved from the Internet <URL:http://www.landesbioscience.com> *

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